紫花苜蓿MsHST基因克隆及拟南芥转化

Cloning of MsHSTGene from Alfalfa and Obtainment of Transgenic Arabidopsis thaliana

运用RT-PCR法从紫花苜蓿(Medicago sativa)中成功克隆出长度约1100bp的尿黑酸法呢基转移酶基因MsHST,采用qRT-PCR的方法,分析MsHST基因的表达特征,结果显示MsHST在所有组织中均有表达,但在叶片中表达量最高,根中最低;随叶片的衰老,表达量降低。且构建了超表达载体pBI-MsHST,并获得拟南芥(Arabidopsis thaliana)转基因植株,经PCR,RT-PCR和GUS染色检测转基因植株,初步证实目的基因在拟南芥中表达。

By using RT-PCR, we cloned a length of 1100 bp of urine black acid base transferase gene from alfalfa （Medicago sativa）. It was named as MsHST, and its expression characteristics were analyzed with qRT-PCR. The results showed that MsHST in all the tested organs, including root, stem, leaf and flower were expressed, but its expression in blade was the highest, expression in root was the lowest. The gene expression was decreased with the senescence of leaf. Using recombinant DNA technology, the MsHST was inserted to expression vector pBI121 to make a construct ofpBI-MsHST. Through Agrobacterium mediated-transfermation, putative transgenic Arabidopsis（Arabidopsis thaliana）plants were obtained, and subjected to PCR, RT-PCR and GUS staining assay, the results showed that the target gene has been integrated to the Arabidopsis thaliana genome and was expressed.